Introduction to OMNISEC system

03/05/2016
Introduction to OMNISEC system
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03/05/2016
OMNISEC
Separations module
OMNISEC RESOLVE
Sample characterization
OMNISEC REVEAL
Introduction to OMNISEC RESOLVE
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OMNISEC RESOLVE
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OMNISEC RESOLVE is the separations module
Integration of all separations components makes
OMNISEC RESOLVE a complete SEC separation
solution
Degasser
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Degassing removes dissolved gases
for better baseline stability and pump
performance
Low-volume (2 x 1 mL channels), high
efficiency degasser
Faster solvent exchange and quicker
equilibration
Better baseline stability and reduced
instrument down-time during solvent
exchange
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SEC pump
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Low pulsation isocratic pump ideal for
SEC separations
 Better resolution
 More repeatable separations
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Integrated seal back-flushing for reduced
seal wear
 Reduced maintenance requirement
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Self-priming pump means no need to
draw mobile phase with a needle, simply
open the purge valve
Autosampler
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Injections from vials or 96-well microtiter plates
Temperature controlled (4-60°C) plate holder
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µl pickup mode
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Zero injection overhead
No sample waste during injections
Partial loop mode
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Preserve protein samples from aggregation
Maintain polymer dissolution
Improve injection of high viscosity samples (e.g. in
DMSO)
45 µl sample overhead
Improved injection accuracy and precision
Post injection washing means undetectable
carry-over
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Column oven
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Store up to 6 analytical
SEC columns or 1 GE
Tricorn 300/10 (e.g.
Superdex) column
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Temperature control from
20 to 65°C
OMNISEC RESOLVE design
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Complete separations module
including columns allows
detector to be plug and play
Two connections to
OMNISEC REVEAL
minimizes setup complexity
Space for solvent and waste
bottles on top
Simply connect to OMNISEC
REVEAL to set up
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Separation module – OMINSEC RESOLVE
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Injection reproducibility 0.2% for maximum result quality
Zero overhead autosampler (vs 50µl in GPCmax)
Temperature controlled autosampler. Cooling to protect samples
from degradation (e.g proteins) or warming reduce sample
viscosity (e.g. samples in DMSO)
Sampling from vials or from 96/384 well-plates
Improved efficiency and smaller volume degasser (2ml vs 30 in
GPCmax – quicker solvent changeover/stabilization)
New pump with integral seal washing
Large capacity column oven with highly effective temperature
control
Room for a vertical GE column in the oven
Introduction to OMNISEC REVEAL
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OMNISEC REVEAL technology
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OMNISEC REVEAL is the integrated detector module
Refractive index
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Refractive index
Measure sample concentration of almost any sample
Measure dn/dc
OMNISEC contains a robust RI cell placing it at front of detector
series
UV/Vis PDA
 Minimizes band broadening (dispersion) between all detectors caused by
RI temperature equilibration tubing
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Wavelength matched with LS detector (640 nm)
RALS/LALS
Intrinsic viscosity
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UV/Vis PDA
Refractive index
UV/Vis PDA
RALS/LALS
Intrinsic viscosity
Light scattering
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Refractive index
The LS detector is a RALS/LALS detector
The laser light is now fibre-delivered
 Better focussing of the laser light
 More power to the cell
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50 mW laser
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Sensitivity improved
UV/Vis PDA
RALS/LALS
Intrinsic viscosity
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Light Scattering
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Refractive index
Molecular weight quantification at 100 ng!
Polystyrene 105 KDa in THF, 0.01 mg/mL, 10 µL injection
Parameter
Retention volume (mL)
Mn (Da)
Mw (Da)
Mz (Da)
PD (Mw/Mn)
100 ng PS105
8.02
105,470
108,180
110,570
1.026
UV/Vis PDA
RALS/LALS
Intrinsic viscosity
OMNISEC REVEAL light scattering sensitivity - Protein
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Molecular weight quantification at 100 ng!
BSA 66 KDa in PBS 0.05 mg/ml, 2 µL injection
Accurate calculation of molecular weight
RV (mL)
Mn (g/mol)
Mw (g/mol)
Mz (g/mol)
Mw/Mn
9.083
63,800
65,070
66,290
1.02
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Viscometer technology
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Refractive index
The exchangeable bridge can be removed and replaced in
just 10 minutes
+IP
SEC IN
UV/Vis PDA
- DP
+
RALS/LALS
OUT
Intrinsic viscosity
Viscometer
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Refractive index
Automated bridge balancing mechanism
As the capillary moves within its sleeve, the resistance changes
altering the bridge balance
UV/Vis PDA
RALS/LALS
Intrinsic viscosity
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Viscometer
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Refractive index
The software automatically searches for the zero position
Viscometer
signal
UV/Vis PDA
0
RALS/LALS
Intrinsic viscosity
OMNISEC REVEAL user-focussed design
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OMNISEC REVEAL is the integrated
detector module
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Developed with the user and
servicing in mind
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OMNISEC REVEAL design
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The user can only interact with those
parts that may need maintenance
Quick and easy replacement of:
 UV bulb
 Viscometer bridge
 Delay columns
Detector module – OMNISEC REVEAL
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Redesigned RALS/LALS with significant sensitivity
improvements
New viscometer design (self-balancing bridge with userexchangeable capillaries)
Improved RI, including change of position in detector
module, and excellent sensitivity
Completely new UV-diode array (full UV/Vis range)
Temperature control of all detectors and inter-detector
tubing (20 to 65°C)
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Systems or detectors
Detector-only or complete system
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Having a complete OMNISEC system
means:
 A complete solution from a single supplier
designed and manufactured to work together
 A single software package for control of the
hardware, data acquisition and data analysis
 A single point of contact for service and support
issues
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Detector-only or complete system
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Taking a detector-only option means:
 Saving money by upgrading an existing
system to advanced detection
 Using an established method and getting
additional information
3rd party
SEC
system
OMNISEC system
AQUEOUS APPLICATIONS
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Conditions for Aqueous analysis
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Eluent:
0.05 M sodium suphate
Flow rate:
1.0 mL/min
Columns:
2 x A6000M
Temperatures °C:
 Autosampler:
 Columns:
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 Detectors: 35
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PEO standards - RI and RALS 196 to 992,000 Da
992,000 Da
196 Da
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PEO 196 Da by Triple Detection !
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PEO 196 Da
3.9 mg/ml, 100 µl injection
Low molecular weight examples
Low dn/dc examples
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Triple Chromatogram of Bisphenol A (MW = 228)
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Mobile Phase: THF
Temperature: 30C
Columns:
Injections:
2 X T2500
100 uL of 3 mg/mL
Flow Rate:
dn/dc:
1 mL/min
0.195
Example Triple Chromatogram of Polypropylene Glycols
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Mobile Phase: THF
Temperature: 30C
Columns:
Injections:
2 X T2500
100 uL of 6 mg/mL
Flow Rate:
dn/dc:
1 mL/min
0.0454
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Summary of Molecular values of Polypropylene Glycols
Samples
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Mw
Mn
PDI
IV
dn/dc
% Recovery
PPG-1A
2,806
2,487
1.13
0.062
0.0454
100.1
PPG-1B
2,644
2,061
1.28
0.061
0.0454
101.6
AVE
2,725
2,274
1.21
0.061
0.0454
100.9
PPG-2A
2,831
2,691
1.05
0.067
0.0454
99.2
PPG-2B
2,753
2,394
1.15
0.061
0.0454
100.1
AVE
2,792
2,543
1.10
0.064
0.0454
99.6
PPG-3A
2,789
2,717
1.03
0.062
0.0454
99.5
PPG-3B
3,217
2,897
1.11
0.071
0.0454
99.4
AVE
3,003
2,807
1.07
0.067
0.0454
99.4
Mobile Phase: THF
Temperature: 30C
Columns:
Injections:
2 X T2500
100 uL of 6 mg/mL
Flow Rate:
dn/dc:
1 mL/min
0.0454
Triple Chromatogram of Polycaprolactone A (Mw = 1,800)
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Mobile Phase: THF
Temperature: 30C
Columns:
Injections:
2 X T2500
100 uL of 8 mg/mL
Flow Rate:
dn/dc:
1 mL/min
0.06
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Triple Chromatogram of Polycaprolactone B (Mw = 1,600)
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Mobile Phase: THF
Temperature: 30C
Columns:
Injections:
2 X T2500
100 uL of 5 mg/mL
Flow Rate:
dn/dc:
1 mL/min
0.06
Example Triple Chromatogram of Silicone Oils (Mw = 44,000)
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Mobile Phase: THF
Temperature: 30C
Columns:
Injections:
2 X T2500
100 uL of 5 mg/mL
Flow Rate:
dn/dc:
1 mL/min
0.029
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OMNISEC REVEAL - summary
Thanks for listening
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